PURIFICATION OF CHYMOTRYPSIN FROM BOVINE PANCREAS USING PRECIPITATION WITH A STRONG ANIONIC POLYELECTROLYTE.
BOERIS, VALERIA; ROMANINI, DIANA; FARRUGIA, BEATRIZ; PICÓ, GUILLERMO
Año: 2009 vol. 44 p. 588 - 592
The separation of chymotrypsin from a crude filtrate of bovine pancreas homogenate was carried out using precipitation with a commercially available negatively charged strong polyelectrolyte: polyvinyl sulfonate. The zymogen form of chymotrypsin was activated by addition of trypsin (0.01 mg/g homogenate), then, the enzyme was precipitated by polyelectrolyte addition at pH 2.5 in the pancreas homogenate. A stoichiometric ratio of 670 bound molecules of chymotrypsin per polyelectrolyte molecule was found in the non-soluble form of the enzymepolyelectrolyte complex. The non-soluble complex was separated by simple centrifugation and re-dissolved by a pH change to 8.0. The recovery of chymotrypsin biological activity was 61% of the initial activity in the homogenate with 4.7-fold increase in its specific activity.